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ANTIBODY & PROTEIN LABELING KITS

Efficient, easy-to-use kits for labeling antibodies and other proteins with our superior CF® Dyes and other labels.
Jump to a section below to choose a kit that’s right for you:

Fast & Simple Antibody Labeling
Easy-to-use with minimal hands-on time, Mix-n-Stain™ kits require no purification after labeling
New IgM Labeling Kits Available
New Mix-n-Stain™ kits for efficient labeling of IgM antibodies with CF® Dyes are now available
Optimal Labeling of Nanobodies®
Mix-n-Stain™ kits specific for labeling Nanobodies® (single variable or VHH domains)
Optimal Labeling for STORM
Your choice of 8 STORM CF® Dyes for labeling antibodies with a low DOL optimal for STORM super-resolution microscopy
Small Ligand Labeling Kits
Create fluorescent SNAP-tag®, CLIP-tag™, Halotag® & other ligands, no purification required
CF® SE Dye Protein Labeling Kits
Larger scale kits, for optimized protein labeling & purification
VivoBrite™ Near-IR Labeling Kits
Kits for labeling antibodies with our superior near-IR CF® Dyes, for in vivo small animal imaging

MIX-N-STAIN™ CF® DYE ANTIBODY LABELING TECHNOLOGY


Mix-n-Stain™ antibody labeling kits dramatically simplify the process of antibody labeling. The resulting conjugates perform comparably to pre-labeled antibodies from leading suppliers. Because the labeling is covalent, the conjugates are stable for long-term storage, and ideal for multi-color imaging and multiplex flow cytometry.

Each Mix-n-Stain™ kit comes with everything you need to perform the conjugation. Without a separation step, you will have an optimally labeled antibody conjugate ready to be used in any immunofluorescence staining experiment.

One breakthrough feature of Mix-n-Stain™ kits is that antibody labeling can be conducted in the presence of many common buffer components such as BSA, gelatin, azide and low concentrations of Tris. As a result, the kits are compatible with most commercial antibody formulations without an antibody purification step prior to the conjugation process.

See below for our selection of Mix-n-Stain™ kits with a variety of label options.

Rapid Antibody Labeling with Next-Generation Fluorescent CF® Dyes, Biotin, & More

With Mix-n-Stain™ CF® Dye Antibody Labeling Kits, you can covalently label as little as ≤5 ug up to 100 ug of an antibody in just 15 minutes, with minimal hands-on time and 100% recovery of antibody. With slight modification in protocol, antibodies can be labeled in the presence of BSA, gelatin, or ascites fluid, for unrivaled convenience. See the Compatibility and Protocol Selection Flowchart below, or download the Product Protocol for a step-by-step worksheet to determine kit compatibility with your antibody.

These kits feature a choice of over 30 superior CF® Dyes, which have advantages in brightness and photostability compared to other commercially available fluorescent dyes. Due to the high water solubility of CF® Dyes, other proteins like BSA or gelatin in the labeling reaction have minimal effect on non-specific background, because non-IgG proteins are washed away during IF staining. We also offer Mix-n-Stain™ kits for biotin, FITC, DIG, and DNP labeling, as well as Mix-n-Stain™ Maxi Antibody Labeling Kits for 1 mg scale labeling reactions.

Features:

  • 3 kit sizes: 5-20 ug, 20-50 ug, or 50-100 ug Ab
  • Choice of over 30 CF® Dye colors, biotin, FITC, or haptens
  • Less than 30 seconds of hands-on time
  • 15 minutes total reaction time
  • No purification of the product is required
  • Compatible with common antibody stabilizers

Compatible with antibody stabilizers and ascites

Monoclonal mouse anti-tubulin clone DM1A in ascites or purified in PBS with 1% BSA was labeled with CF®488A Mix-n-Stain™ using the modified protocol for total protein labeling. The Mix-n-Stain™-labeled antibodies performed comparably in immunofluorescence staining of HeLa cells as unlabeled antibody (in ascites) detected with secondary antibody.

Combine direct & indirect IF for flexible multiplexing

Combined direct and indirect immunofluorescence staining of rat testis with mouse anti-tubulin and CF®488A goat anti-mouse (min x rat) (microtubules, green), CF®555 Mix-n-Stain™ labeled mouse anti-ZO1 (tight junctions, red) and CF®640R phalloidin (actin filaments, cyan). See our Tech Tip: Combined Direct & Indirect Immunofluorescence Using Primary Antibodies from the Same Host.

Featuring CF® Dyes with superior brightness

Mouse anti-transferrin receptor antibody from (endosome and plasma membrane marker) was labeled using Lightning-Link® Rapid DyLight® Conjugation Kits from Novus Biologicals (left) or Mix-n-Stain™ CF® Dye Antibody Labeling Kits (right) according to manufacturers’ instructions. The CF® Dye conjugates show higher signal and more specific staining compared the DyLight®conjugates when imaged using the same settings, due to the superior brightness of CF® Dyes. The insets show the same field of view imaged with a higher gain setting.

Mix-n-Stain™ CF® Dye, Biotin, FITC & Hapten IgG Antibody Labeling Kits

Product description Label options Labeling time
Mix-n-Stain™ CF® Dye Antibody Labeling Kits Choice of over 30 bright & photostable CF® Dyes 15 minutes (~30 seconds hands-on time)

Download the product protocol
Mix-n-Stain™ Cyanine Dye
Antibody Labeling Kits		 Cyanine 555
Cyanine 647
Mix-n-Stain™ FITC Antibody Labeling Kits FITC
Mix-n-Stain™ Biotin Antibody Labeling Kits Biotin
Mix-n-Stain™ DNP Antibody Labeling Kits DNP
Mix-n-Stain™ Digoxigenin Antibody Labeling Kits Digoxigenin (DIG)
Mix-n-Stain™ Maxi Antibody Labeling Kits Choice of 13 bright & photostable CF® Dyes, FITC, Cyanine 555, and Cyanine 647 30 minutes (~30 seconds hands-on time)
Mix-n-Stain™ STORM CF® Antibody Labeling Kits Choice of 8 CF® Dyes specialized for STORM super-resolution microscopy

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MIX-N-STAIN™ CF® DYE IGM ANTIBODY LABELING KITS

Finally! Simple & Efficient Labeling of IgM Antibodies

IgM antibodies are difficult to label with fluorescent dyes because the conditions typically used for labeling IgG generally do not result in bright conjugates with good antibody reactivity for IgM. Biotium has resolved this issue by optimizing the reaction conditions for labeling IgM antibodies with Mix-n-Stain™ CF® Dye IgM Antibody Labeling Kits.

Features

  • Label 25 ug or 100 ug of IgM antibody
  • Choice of 8 CF® Dye colors and FITC
  • 15-30 minute total reaction time
  • No purification, 100% recovery

 

Mouse anti-beta tubulin clone 5H1 (IgM kappa) was labeled with CF®488A using Mix-n-Stain™ IgM labeling, then was used to stain methanol-fixed HeLa cells at 5 ug/mL (right panel). The conjugate shows comparable staining specificity compared to unlabeled antibody used at 1 ug/mL and detected using CF®488A Goat Anti-Mouse IgM Mu chain secondary antibody at 2 ug/mL (left panel).

Mix-n-Stain™ CF® Dye IgM Antibody Labeling Kits

Conjugation Ex/Em Size Catalog No. Dye Features
CF®405L 413/547 nm 25 ug 92558 CF®405L Features
100 ug 92559
CF®405M 416/452 nm 25 ug 92560 CF®405M Features
100 ug 92561
CF®488A 490/516 nm 25 ug 92562 CF®488A Features
100 ug 92563
CF®555 554/568 nm 25 ug 92564 CF®555 Features
100 ug 92565
CF®568 562/583 nm 25 ug 92566 CF®568 Features
100 ug 92567
CF®594 593/615 nm 25 ug 92568 CF®594 Features
100 ug 92569
CF®640R 642/663 nm 25 ug 92570 CF®640R Features
100 ug 92571
CF®647 652/668 nm 25 ug 92572 CF®647 Features
100 ug 92573
FITC 498/517 nm 25 ug 92574
100 ug 92575
Note: Before selecting a kit, please download the Product Protocol to see Kit Compatibility and Protocol Selection guidelines.

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MIX-N-STAIN™ NANOBODY LABELING KITS


Mix-n-Stain™ Nanobody Labeling Kits are designed specifically for labeling single-domain Nanobodies® (also called camelid single variable or VHH domains) with one of Biotium’s bright and photostable CF® Dyes or biotin. The kits allow optimal labeling of between 5-50 ug of Nanobody® in just 30 minutes, with minimal hands-on time and no purification. Labeling tolerates common buffer additives, including BSA.

The kits come with a selection of our next-generation CF® Dyes or biotin. CF® Dyes have advantages in brightness and photostability compared to other commercially available fluorescent dyes (learn more about CF® Dyes).

Features

  • Label 5-20 ug or 20-50 ug of Nanobody®
  • Choice of 7 CF® Dye colors or biotin
  • Simple 30 minute labeling, no purification
  • Compatible with common stabilizers
  • Dye options for super-resolution and 2-photon imaging (learn more)
Anti-histone Nanobody® was labeled using the Mix-n-Stain™ CF®488A 5-20 ug Nanobody Labeling Kit, then used to stain methanol-fixed HeLa cells at 2 ug/mL. Labeling either 5 ug or 20 ug of Nanobody® generated bright and specific conjugates, but protein amounts at the upper end of the kit range were more optimal. Adding 15 ug BSA to 5 ug Nanobody® to bring the protein amount to 20 ug produced more optimal labeling and brighter staining. Labeled BSA did not affect specificity of staining.

Overview of the Mix-n-Stain™ Nanobody Labeling Protocol.

Mix-n-Stain™ Nanobody Labeling Kits

Label Ex/Em Labeling size
(ug Nanobody®)		 Catalog number
Biotin N/A 5-20 ug 92500
20-50 ug 92501
CF®405S 404/431 nm 5-20 ug 92502
20-50 ug 92503
CF®488A 490/515 nm 5-20 ug 92504
20-50 ug 92505
CF®568 562/583 nm 5-20 ug 92506
20-50 ug 92507
CF®594 593/614 nm 5-20 ug 92508
20-50 ug 92509
CF®640R 642/662 nm 5-20 ug 92510
20-50 ug 92511
CF®647 650/665 nm 5-20 ug 92512
20-50 ug 92513
CF®680R 680/701 nm 5-20 ug 92514
20-50 ug 92515
Note: Before selecting a kit, please download the Product Protocol to see Kit Compatibility and Protocol Selection guidelines.

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MIX-N-STAIN™ STORM CF® DYE ANTIBODY LABELING KITS

Mix-n-Stain™ STORM CF® Dye Antibody Labeling Kits offer a choice of 8 CF® Dyes developed with superior brightness and unique photoswitching properties that are ideal for super-resolution imaging by STORM. In addition, these specialized antibody labeling kits are designed to offer a low degree of labeling (DOL, or dye molecules per antibody molecule) of 1-2.5, which is reported to be optimal for STORM (Bittel et al. 2015). Biotium has also developed novel green-excited STORM dyes CF®583R and CF®597R to expand the color palette for multi-color STORM. Learn more about the UC Berkeley and Biotium collaboration paper on CF®583R and CF®597R dyes for STORM.

Features

  • Choice of 8 CF® Dye colors ideal for STORM
  • CF®583R and CF®597R are novel green-excited STORM dyes
  • Labeling optimized to provide low 1-2.5 DOL
  • Less than 30 seconds of hands-on time
  • 30 minutes total reaction time
  • No purification, 100% recovery

Learn more about CF® Dyes for STORM or our other fluorescent reagents validated for super-resolution microscopy.

Typical (d)STORM image of the actin cytoskeleton in a fixed COS-7 cell labeled by phalloidin-CF®583R. (d)STORM was performed under standard conditions using a standard tris-based (d)STORM imaging buffer containing 100 mM cysteamine and an oxygen scavenger. Image courtesy of Bowen Wang, Michael Xiong, and Professor Ke Xu, College of Chemistry, University of California, Berkeley.

Two-color (d)STORM imaging of a fixed COS-7 cell: microtubules were immunolabeled with CF®647 conjugate, and mitochondria were immunolabeled with CF®583R conjugate. (d)STORM was performed under standard conditions using a standard tris-based (d)STORM imaging buffer containing 100 mM cysteamine and an oxygen scavenger. CF®647 was first imaged with 647 nm excitation, and CF®583R was next imaged with 561 nm excitation. Images courtesy of Bowen Wang, Michael Xiong, and Professor Ke Xu, College of Chemistry, University of California, Berkeley.

Mix-n-Stain™ STORM CF® Dye Antibody Labeling Kits

Conjugation Ex/Em Size Catalog No. Dye Features
CF®505 505/519 nm 50 ug 92549
CF®568 562/584 nm 50 ug 92550 CF®568 Features
CF®583R 585/609 nm 50 ug 92551 CF®583R Features
CF®597R 597/619 nm 50 ug 92553 CF®597R Features
CF®647 652/668 nm 50 ug 92554 CF®647 Features
CF®660C 667/685 nm 50 ug 92555 CF®660C Features
CF®680 681/698 nm 50 ug 92556 CF®680 Features
CF®750 755/779 nm 50 ug 92557 CF®750 Features

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MIX-N-STAIN™ CF® DYE SMALL LIGAND LABELING KITS


Mix-n-Stain™ CF® Dye small ligand labeling kits are designed for rapid labeling of small (MW ~ 150 – 5,000) and relatively high affinity biological ligands (or substrates). Labeling takes about 30 minutes, without a final purification step. The ligands to be labeled must contain an aliphatic amine group that is not required for biological activity of the ligand. The amine group will form a covalent linkage with the reactive CF® Dye provided in the kit. For example, suitable ligands or substrates include SNAP-tag®, CLIP-tag™ and HaloTag® ligands with an aliphatic amine. Many other small ligands are also possible candidates if they meet the criteria described above.

Simply mix your small molecule ligand with the reaction buffer and the optimally formulated dye provided, and incubate for 30 minutes, followed by a brief 5 minute quenching step. No reactive Mix-n-Stain™ dye is available at the end of labeling; therefore the conjugate is ready for cell staining and imaging without further purification. Mix-n-Stain™ labeled ligands perform comparably to commercially-available pre-labeled ligands (Figure 1), at a fraction of the cost.

For live cell imaging, the Mix-n-Stain™ small ligand kits are ideal to generate fluorescent chemical tags that provide smaller, brighter, and more versatile alternatives to fluorescent proteins. Labeling of various chemical tags with Mix-n-Stain™ kits yielded highly efficient and specific labeling of the target proteins both on surface and inside of living cells. The Mix-n-Stain™ small ligand kits have also been proven compatible with fixed cell imaging and gel-based assays.

Features:

  • Choice of 10 CF® Dye colors for extracellular staining
  • Choice of 5 CF® Dye colors for intracellular staining
  • For labeling molecules with a MW of 150-5000, containing an inactive aliphatic amine
  • Suitable ligands or substrates include SNAP-tag®, CLIP-tag™ and HaloTag® ligands
  • 30 minutes total reaction time, no purification necessary
Figure 1. Live cell imaging of HeLa cells expressing CLIP-NK1R labeled with (A) CLIP-tag™ ligand conjugated to green fluorescent CF®488A using Mix-n-Stain™ compared to (B) CLIP-surface 488 from New England BioLabs. Cell nuclei were stained blue with Hoechst 33342.
Figure 2. Three color imaging in live HeLa cells. Cell surface protein ADRβ2 was labeled using the CF®488A Mix-n-Stain™ kit via the SNAP-tag® (green). Mitochondrial protein Cox8A was labeled using the CF®540 Mix-n-Stain™ kit via the CLIP-tag™ (red). Cell nuclei were stained blue with Hoechst 33342.

Mix-n-Stain™ Small Ligand Kits

Product description Label options Labeling time
Mix-n-Stain™ CF® Dye Small Molecule Labeling Kits Extracellular Staining: Choice of 10 bright and stable CF® Dyes ~30 minutes (minimal hands-on time)

Download the product protocol
Intracellular Staining: Choice of 5 bright and stable CF® Dyes

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CF® DYE SE
PROTEIN LABELING KITS

CF® Dye SE Protein Labeling Kits allow you to label your antibody of interest with one of our many bright and photostable CF® Dyes. The difference is that while the Mix-n-Stain™ kits come optimized for consistent labeling of small amounts of antibody, the SE Protein Labeling Kits are designed for 3 labelings of up to 1 mg protein with removal of free dye after labeling. The SE Protein Labeling Kits allow the user to optimize the amount of dye and antibody and determine the degree of protein labeling, for optimization of labeling and antibody performance.

Features:

  • Choice of 19 CF® Dye colors or biotin
  • Reagents for 3 labeling reactions with purification
  • Each reaction can label up to 1 mg of antibody or protein
  • Includes instructions for determining degree of labeling

VIVOBRITE™ NEAR-IR ANTIBODY LABELING KITS FOR IN VIVO IMAGING

VivoBrite™ Antibody Labeling Kits use the same labeling technology as our SE Protein Labeling Kits. The kits include the same components as the SE Protein Labeling Kits, with the addition of a syringe and syringe filter to sterile filter the labeled protein, allowing it to be used for in vivo small animal studies. The VivoBrite™ kits are offered in 4 near-IR CF® Dye colors, which are useful for small animal in vivo imaging. Biotium’s near-IR fluroescent dyes are best in class for brightness, photostability, and water solubility.

Features:

  • Choice of 4 near-IR CF® Dye colors
  • Kit contains reagents for 3 labeling reactions
  • Each reaction can label up to 1 mg of antibody
  • Kit can also be used to label other proteins
  • The resulting labeled protein is purified and sterile

Protein Labeling Kits

Product description Label options Product protocol
CF® Dye SE Protein Labeling Kits 19 CF® Dye colors across the spectrum, plus biotin PI-CF® Dye SE Protein Labeling Kits
VivoBrite™ Antibody Labeling Kits for Small Animal In Vivo Imaging 4 near-IR CF® Dyes PI-VivoBrite™ Antibody Labeling Kits

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SNAP-tag is a registered trademark of New England Biolabs; CLIP-tag is a trademark of New England Biolabs; HaloTag is a registered trademark of Promega; Nanobody is a registered trademark of Ablynx.

Associated products

Mix-n-Stain™ Biotin Antibody Labeling Kit

From: $129 Sizes: 3x(5-20 ug) labelings, 5-20 ug labeling, 20-50 ug labeling, 50-100 ug labelingCatalog #:
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Mix-n-Stain™ CF® Dye Antibody Labeling Kits

From: $129 Sizes: 3x(5-20 ug) labelings, 5-20 ug labeling, 20-50 ug labeling, 50-100 ug labelingCatalog #:
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Mix-n-Stain™ CF® Dye Small Ligand Labeling Kits

From: $206 Sizes: 1 labelingCatalog #:
92350, 92351, 92352, 92353, 92354, 92355, 92356, 92357, 92358, 92359, 92360, 92361, 92362, 92363, - 92364View allHide
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Mix-n-Stain™ Digoxigenin Antibody Labeling Kit

From: $141 Sizes: 3x(5-20 ug) labelings, 5-20 ug labeling, 20-50 ug labeling, 50-100 ug labelingCatalog #:
92450, 92328, 92329, - 92330View allHide
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Mix-n-Stain™ FITC Antibody Labeling Kit

From: $81 Sizes: 5-20 ug labeling, 20-50 ug labeling, 50-100 ug labelingCatalog #:
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CF® Dye & Biotin SE Protein Labeling Kits

From: $497 Sizes: 3 labelingsCatalog #:
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VivoBrite™ Rapid Antibody Labeling Kits for Small Animal In Vivo Imaging

From: $536 Sizes: 3 labelingsCatalog #:
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Mix-n-Stain™ Maxi Antibody Labeling Kits, 1 mg Labeling

From: $597 Sizes: 1 mg labelingCatalog #:
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Mix-n-Stain™ Nanobody Labeling Kits

From: $144 Sizes: 5-20 ug labeling, 20-50 ug labelingCatalog #:
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Faq

Mix-n-Stain™ Antibody Labeling Kits
My product was accidentally left out at room temperature or exposed to light. Is it ruined?
Most of our products are stable at room temperature for many days, so in all likelihood the product will still work just fine. To be on the safe side, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples. One exception that we are aware of is GelGreen™, which is more sensitive to light exposure than most of our other fluorescent dyes. If GelGreen™ is exposed to ambient light for a prolonged period of time (days to weeks), its color will change from dark orange to brick red. If this occurs, the GelGreen will no longer work for gel staining.  


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Where can I find the expiration date or shelf life of a product? Is the product still good if I have had the product longer than the guaranteed shelf life?
Bioscience kits The guaranteed shelf life from date of receipt for bioscience kits is listed on the product information sheet. Some kits have an expiration date printed on the kit box label, this is the guaranteed shelf life date calculated from the day that the product shipped from our facility. Kits often are functional for significantly longer than the guaranteed shelf life. If you have an older kit in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the kit still works for your application before processing a large number of samples or precious samples. Antibodies and other conjugates The guaranteed shelf life from date of receipt for antibodies and conjugates is listed on the product information sheet. Antibodies and other conjugates often are functional for significantly longer than the guaranteed shelf life. If you have an older conjugate in storage that you wish to use, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples. For lyophilized antibodies, we recommend reconstituting the antibody with glycerol and antimicrobial preservative like sodium azide for the longest shelf life (note that sodium azide is not compatible with HRP-conjugates). Chemicals, dyes, and gel stains Biotium guarantees the stability of chemicals, dyes, and gel stains for at least a year from the date you receive the product. However, the majority of these products are highly stable for many years, as long as they are stored as recommended. Storage conditions can be found on the product information sheet or product safety and data sheet, material safety data sheet, and on the product label. Fluorescent compounds should be protected from light for long term storage. If you have a Biotium compound that has been in storage for longer than one year that you wish to use, we recommend performing a small scale positive control experiment to confirm that the compound still works for your application before processing a large number of samples or precious samples. Expiration date based on date of manufacture (DOM) If your institution requires you to document expiration date based on date of manufacture for reagents, please contact techsupport@biotium.com for assistance. Chemical products with special stability considerations: Esters Ester compounds include the following: • Succinimidyl esters (SE, also known as NHS esters), such as our amine-reactive dyes • Acetoxymethyl esters (AM esters) such as our membrane-permeable ion indicator dyes • Diacetate-modified dyes, like ViaFluor™ 405, CFDA, and CFDA-SE cell viability/cell proliferation dyes Ester dyes are stable in solid form as long as they are protected from light and moisture. Esters are not stable in aqueous solution. Concentrated stock solutions should be prepared in anhydrous DMSO (see Biotium catalog no. 90082). Stock solutions in anhydrous DMSO can be stored desiccated at -20°C for one month or longer. Esters should be diluted in aqueous solution immediately before use. Succinimidyl esters (SE) should be dissolved in a solution that is free of amine-containing compounds like Tris, glycine, or protein, which will react with the SE functional group. AM esters and diacetate compounds should be dissolved in a solution that is free of serum, because serum could contain esterases that would hydrolyze the compound. A note on CF® Dye succinimidyl ester stability Succinimidyl esters (SE) are generally susceptible to hydrolysis, which can result in lower labeling efficiency. Many commercially available fluorescent dyes used for life science research are heavily sulfonated dyes which makes them particularly hygroscopic, worsening the hydrolysis problem. In addition, for several commercially available SE reactive dyes, the SE group is derived from an aromatic carboxylic acid, while the SE group in all of Biotium’s CF® Dyes is prepared from an aliphatic carboxylic acid. This structural difference reduces the susceptibility of CF® Dye SE reactive groups to hydrolysis, resulting in relatively stable reactive dyes with consistently higher labeling efficiency compared to other SE derivatives of other fluorescent dyes. Maleimides, MTS and thiosulfate dyes Like the succinimidyl ester dyes, these dyes are also susceptible to hydrolysis, although generally to a much lower degree. Thus, for long term storage, anhydrous DMSO is recommended for making stock solutions. Other reactive dyes Amines, aminooxy (also known as oxylamine), hydrazide, azide, alkyne, BCN, and tyramide reactive dyes, as well as dye free acids, are generally stable in aqueous solution when stored at -20°C for 6-12 months or longer, as long as no compounds are present that may react with the dye’s functional group. See the product information sheets for specific reactive dyes more information. Coelenterazines and D-luciferin Coelenterazines are stable in solid form when stored as recommended; they are not stable in aqueous solution. Concentrated coelenterazine stock solutions (typically 1-100 mg/mL) should be prepared in ethanol or methanol; do not use DMSO or DMF to dissolve coelenterazines, because these solvents will oxidize the compounds. Ethanol or methanol stocks of coelenterazine can be stored at -20°C or below for six months or longer; alcohol stocks may evaporate during storage, so use tightly sealing screw cap vials and wrap the vials with Parafilm for long term storage. Propylene glycol also can be used as a solvent to minimize evaporation. If the solvent evaporates, the coelenterazine will still be present in the vial, so note the volume in the vial prior to storage so that you can adjust the solvent volume to correct for evaporation if needed. Prepare working solutions in aqueous buffers immediately before use. Coelenterazines are stable for up to five hours in aqueous solution. Aquaphile™ coelenterazines are water soluble formulations of coelenterazines. They are stable in solid form when stored as recommended. Aquaphile™ coelenterazines should be dissolved in aqueous solution immediately before use. They are stable for up to five hours in aqueous solution. Note that coelenterazines are predominantly yellow solids, but may contain dark red or brown flecks. This does not affect product stability or performance. If your coelenterazine is uniformly brown, then it is oxidized and needs to be replaced. D-luciferin is stable in solid form and as a concentrated stock solution when stored as recommended; it is not stable at dilute working concentrations in aqueous solution. Prepare concentrated D-luciferin stock solutions (typically 1-100 mg/mL) in water, and store in aliquots at -20°C or below for six months or longer. Prepare working solutions immediately before use.


View more FAQs
Where can I find technical details for all of the CF® dyes?
See our CF® Dye Quick Reference Table for a list of dyes and summary of their features. Our CF® Dye Selection Guide has more detailed information on each CF® dye, and ordering information for our various CF® dye product lines. You can download CF® dye normalized absorption and emission data in Excel format.


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CF® Dyes
Do you offer alternatives to Akoya Opal™ tyramides?
See below for our recommended alternatives to Akoya tyramides. Unless noted as a direct replacement, these tyramide dyes are spectrally similar alternatives. They have not been validated as direct replacements for use with Akoya kits or imaging systems.

CF® Dye Alternatives for Akoya Tyramides

Akoya Tyramide Ex/Em (nm) Biotium Tyramide Ex/Em (nm) Biotium Cat. No. Notes CF® Dye Features
TSA Coumarin 402/443 CF®405S 404/431 92197 Much brighter & more
photostable alternative		 CF®405S Features
Opal™ Polaris 480 450/500 CF®430 426/498 96053 Recommended
for abundant targets		 CF®430 Features
Opal™520 494/525 CF®488A 490/515 92171 CF®488A Features
TSA Fluorescein 494/517 Fluorescein 492/514 96018 Direct replacement See CF®488A for a brighter &
much more photostable alternative
TSA TMR
TSA Cyanine 3		 550/570 Cyanine 555 555/565 96020 Direct replacement
for Cyanine 3		 See CF®555 & CF®568 for brighter
& more photostable alternatives.
CF®555 555/565 96021 Brighter & more
photostable alternatives		 CF®555 Features
CF®568 562/583 92173 CF®568 Features
Opal™570 550/570 CF®550R 551/577 96077 CF®550R Features
CF®555 555/565
 96021 CF®555 Features
TSA Plus Cyanine 3.5 581/596 CF®583R 586/609 96085 Brighter & more
photostable alternatives		 CF®583R Features
CF®594 593/614 92174 CF®594 Features
Opal™620 588/616 CF®583R 586/609 96085 CF®583R Features
TSA Cyanine 5 648/667 CF®640R 642/662 92175 Brighter & more
photostable alternative		 CF®640R Features
CF®647 650/665 96022 Brighter alternative CF®647 Features
CF®660R 663/682 92195 Less cross-talk with visible red dyes;
bright & extremely photostable		 CF®660R Features
TSA Plus Cyanine 5.5
Opal™690		 673/692
676/694		 CF®680R 680/701 92196 Bright & extremely
photostable		 CF®680R Features
Opal™ Polaris 780 750/770 CF®754 745/786 96090 Recommended
for abundant targets		 Unique NIR dye for TSA
TSA Biotin N/A Biotin-XX N/A 92176 Direct replacement
TSA Plus DNP N/A DNP N/A 96019 Direct replacement
TSA Plus DIG N/A N/A N/A N/A N/A
Note: The CF® Dyes listed here are spectrally similar alternatives to Opal™ dyes, they have not been validated as direct replacements for use with Akoya's kits or imaging system. Opal is a trademark of Akoya Biosciences.  


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My product was accidentally left out at room temperature or exposed to light. Is it ruined?
Most of our products are stable at room temperature for many days, so in all likelihood the product will still work just fine. To be on the safe side, we recommend performing a small scale positive control experiment to confirm that the product still works for your application before processing a large number of samples or precious samples. One exception that we are aware of is GelGreen™, which is more sensitive to light exposure than most of our other fluorescent dyes. If GelGreen™ is exposed to ambient light for a prolonged period of time (days to weeks), its color will change from dark orange to brick red. If this occurs, the GelGreen will no longer work for gel staining.  


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Which CF® Dye Bungarotoxin will have the lowest background?
Dyes that carry multiple negative charges can introduce background. Usually, this is more of a concern with labeled antibodies that carry many dyes, as opposed to a small toxin like bungarotoxin. When staining tissues, the endogenous autofluorescence of the tissue itself is often the most significant source of background. Endogenous fluorescence background in tissue is usually highest in the blue wavelengths (DAPI channel) and lowest in the far-red (Cy®5 channel). Our CF®633 bungarotoxin (catalog no. 00009) is a far-red conjugate for the Cy®5 channel with a low negative charge that should have low background from either the dye or autofluorescence. We test fluorescent bungarotoxin on rat skeletal muscle sections. While the tissue shows autofluorescence, the bungarotoxin staining of motor endplates is usually much brighter than the background for all of the dye colors we’ve tested.  However, if you are staining human tissue (especially brain), lipofuscin autofluorescence may be bright in all channels. This usually shows up as bright, punctate dots around cell nuclei. While we would usually recommend our TrueBlack® lipofuscin quenchers for human brain tissue, they are not compatible with bungarotoxin staining. We have, however, found that EverBrite TrueBlack® Mounting Medium (cat. no. 23017) can be used to mount skeletal muscle sections stained with bungarotoxin. Cy Dye is a registered trademark of Cytiva.


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